Compositions and methods for capturing, isolating, detecting, analyzing and quantifying macromolecules

Abstract

A method for making an imprint composition capable of capturing a macromolecule of interest, comprising the steps of (a) solidifying a fluidic matrix material capable of retaining shaped cavities when in a solid or semi-solid form in the presence of a template molecule that corresponds to a portion of the macromolecule, thereby yielding a solid or semi-solid matrix material having template molecules entrapped therein; and (b) removing the template molecules from the solid or semi-solid matrix material, yielding an imprint composition capable of capturing the macromolecule.

What is claimed is:  
     
       1. A method of making an imprint composition capable of capturing a macromolecule of interest, comprising the steps of: 
       (a) solidifying a fluidic matrix material capable of retaining shaped cavities when in a solid or semi-solid form in the presence of a template molecule that corresponds to a portion of the macromolecule, thereby yielding a solid or semi-solid matrix material having template molecules entrapped therein; and  
       (b) removing the template molecules from the solid or semi-solid matrix material, yielding an imprint composition capable of capturing the macromolecule.  
     
     
       2. The method of  claim 1  in which the fluidic matrix material is a heat-sensitive composition and the solidifying step comprises lowering the temperature of the heat-sensitive composition from above its melting point to below its melting point. 
     
     
       3. The method of  claim 2  in which the heat-sensitive composition is selected from the group consisting of hydrogels, agarose, gelatins and moldable plastics. 
     
     
       4. The method of  claim 1  in which the fluidic matrix material is a composition comprising a polymerizable compound and the solidifying step comprises polymerizing the polymerizable compound. 
     
     
       5. The method of  claim 4  wherein the polymerizable compound is selected from the group consisting of styrene, methyl methacrylate, 2-hydroxyethyl methacrylate, 2-hydroxyethyl acrylate, methyl acrylate, acrylamide, vinyl ether, vinyl acetate, divinylbenzene, ethylene glycol dimethacrylate, ethylene glycol diacrylate, pentaerythritol dimethacrylate, pentaerythritol diacrylate, N,N′-methylenebisacrylamide, N,N′-ethylenebisacrylamide, N,N′-(1,2-dihydroxyethylene)bis-acrylamide and trimethylolpropane trimethacrylate. 
     
     
       6. The method of  claim 1  in which the template molecule is immobilized on a solid support or substrate. 
     
     
       7. The method of  claim 1  which the solidifying step is carried out under conditions which are denaturing with respect to the template molecule. 
     
     
       8. The method of  claim 1  in which the solidifying step is carried out under conditions which are native with respect to the template molecule. 
     
     
       9. The method of  claim 1 , wherein the template molecule corresponds to a terminal portion of the macromolecule. 
     
     
       10. The method of  claim 1 , wherein the macromolecule is a polynucleotide and the template molecule comprises an oligonucleotide. 
     
     
       11. The method of  claim 1 , wherein the macromolecule is a polypeptide and the template molecule comprises an oligosaccharide or a glycopeptide. 
     
     
       12. The method of  claim 1 , wherein the macromolecule is a polypeptide and the template molecule comprises a peptide. 
     
     
       13. The method of  claim 12 , wherein the sequence of the peptide corresponds to a contiguous sequence of the polypeptide. 
     
     
       14. The method of  claim 13 , wherein the peptide is between 3 and 30 amino acids in length. 
     
     
       15. The method of  claim 13 , wherein the peptide is between 4 and 15 amino acids in length. 
     
     
       16. The method of  claim 13 , wherein the peptide is between 4 and 7 amino acids in length. 
     
     
       17. The method of  claim 13 , wherein the portion of the polypeptide comprises the C-terminus of the polypeptide, the N-terminus of the polypeptide or an internal sequence of the polypeptide.